Aberrant expression of ARE-binding proteins (ARE-BPs) plays an important role in several diseases, including cancer. Both tristetraprolin (TTP) and human antigen R (HuR) are important ARE-BPs and always play opposite roles in regulating target mRNAs. Our previous work has demonstrated that TTP expression is decreased in gastric cancer (GC). In this study, we reported that HuR was elevated in GC cell lines and gastric cancer patients and that decreased TTP expression partly contributed to the elevated HuR levels by regulating its mRNA turnover. We also observed that dysregulation of TTP and HuR elevated the high-mobility group box 1 (HMGB1) expression in different ways. HuR promoted HMGB1 expression at translational level, while TTP regulated HMGB1 mRNA turnover by destabilizing its mRNA. Increased HuR promoted cancer cell proliferation and the metastasis potential partly by HMGB1. Using immunohistochemistry, we observed that both positive cytoplasmic and high-expression of nuclear HuR were associated with poor pathologic features and survival of GC patients. In conclusion, this study demonstrated that dysregulation of the TTP and HuR plays an important role in GC. Moreover, high HuR nuclear expression or aberrant cytoplasmic distribution may serve as a predictor of poor survival.